DNA Courses

FloCyte has several possible programs for DNA studies, a Multiparameter DNA Cell Cycle module, with Cell specific Proteins / cytokines, and Apoptosis. These can be coupled with a software training program on Modfit from Verity.

Basics of Cell Cycle Analysis

A very brief history of cell cycle

Going from cell cycle to single parameter DNA histogram

What do DNA histograms tell us – what other information is required for interpretation?

Brief discussion of DNA dyes – do’s and don’ts

Examples of perturbations in DNA histograms

Sub-G1 cells – how to identify apoptotic cells from debris

Multiparameter Cell Cycle

BrdU to more precisely determine S phase

Traditional method

SBIP method

Other assays for proliferation – Ki-67, cyclins

Early two parameter approaches

DNA versus protein

DNA versus RNA

Native versus Denatured DNA for identification of mitosis

Use of cyclins to study cell cycle

Background on role of cyclins, cdks and ckis

Discussion of each of the useful cyclins: D, E, A, and B

Use of cyclins to study:

Refinement of cell cycle compartments

Drug treatment

Unbalanced growth

Drug point of action – stathmokinesis

Difference between cancer and normal cells

Three parameter approaches using cyclins and DNA

Multiparameter approaches using modification of chromatin proteins

Phosphorylation of histone H3 to identify mitotic cells

Phosphorylation of histone H2AX to identify DNA double-strand breaks

Sensitivity of the assay

Detection of DNA damage by a variety of clastogens

Measuring DNA damage using ATM

Other multiparameter approaches

Inactivated versus total retinoblastoma protein

Our apoptosis curriculum covers the basic understanding of what is happening in programmed cell death vs necrosis, and how to measure this using flow Cytometry as a tool. Topics covered include

Tunel Assays

Caspases and how they relate to flow, and the assays which measure them, DEVD-Rhod110 and D2R for example